M32 – Zeiss Z1 Lightsheet Microscope

System Description

Lightsheet microscopy is the method of choice for delicate living samples and for large cleared samples. A sheet-shaped light beam is used for exciting a single plane of the sample. The fluorescent light from the excited plane is imaged by perpendicular, camera based detection. This results in optical sectioning with low-dosage excitation and next to no phototoxic stress for living samples. Only in-focus volumes that are actually imaged, are illuminated. Using long-distance objectives and fast z-scanning, large cleared samples (up to 1x1x1 cm) can be acquired in short time.

The M32 Zeiss Z1https://www.zeiss.com/microscopy/int/light/lightsheet-z-1.html lightsheet microscope allows for bidirectional excitation, sample rotation, tile scanning, z-scanning and time-lapse imaging. Dual cameras provide fast acquisition for two channels, although with intermittent filter changes, more colors can be acquired easily. An optical zoom ranges from 0.36x to 2.5 x, although reasonable beam shapes (edges not thicker than two times the beam waist thickness) require a zoom of at least 0.71. Samples are mounted on a vertical, hanging support, dipped into the medium and illuminated and imaged horizontally. Scanning perpendicular to the light sheet plane is done by moving the sample through the light sheet. 3D scans are limited to a maximum speed of one stack per second.

The best achievable resolving power along the z-axis of the detection objective is about 2µm (20x/1.0 objective at optical zoom 2.5), providing cellular and sometimes sub-cellular resolution. For cleared samples, proper refractive index matching of embedding medium and imaging medium to the objective is of paramount importance. See the list of available objectives below.

The system is able to run multidimensional experiments including any combination of the following:

Illumination

White Light LED

Laser for Lightsheet

405nm 20mW 488nm 50mW 561nm 50mW 638nm 60mW

Beamsplitter & Filters

The 5 Beamsplitters divide the emission light into two channels, each of which is detected by a PCO edge sCMOS camera. The beamsplitters are associated with fixed emission filters, for camera one and camera two, respectively.